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Activation of Phospholipid Metabolism During Mediator Release from Stimulated Rat Mast Cells¹

From the Department of Medicine, Washington University School of Medicine, 660 South Euclid Avenue, St. Louis, Missouri 63110

Abstract

Phospholipid metabolism was studied during mediator release from highly purified rat mast cells. The incorporation of ³²PO₄ into individual phospholipids was determined after isolation of phospholipid classes by two-dimensional thin layer chromatography. Various stimulators of histamine release (anti-IgE antibody, concanavalin A [Con A], compound 48/80, or the ionophore A23187) increased ³²PO₄ incorporation into phosphatidic acid (PA), phosphatidylinositol (PI), and phosphatidylcholine (PC) 4- to 10-fold within 15 min. No change in ³²PO₄ incorporation into phosphatidylserine (PS), phosphatidylethanolamine, or sphingomyelin was detected. Anti-IgE antibody caused significantly increased labeling of PA within 8 sec and of PI and PC within 30 sec. The concentrations of anti-IgE and Con A causing threshold, half maximal, and maximal phospholipid labeling were slightly less than those required for comparable changes in histamine release. Both phospholipid labeling and mediator release in response to anti-IgE or Con A were enhanced 3- to 4-fold by PS. The addition of 50 mM -methylmannoside to mast cells 10 min after challenge with Con A rapidly halted both ongoing mediator release and PA labeling. The results of these studies indicate that marked and selective changes in phospholipid metabolism occur before as well as during mediator release from mast cells and that these reactions may be an intrinsic part of the biochemical mechanisms that control mediator release.

Footnotes

¹ This work was supported by National Institutes of Health Grants NIAID 1 RO AI 13380 and 1 PO1 AI 12450.

² Donald Kennerly is a doctoral candidate at Washington University School of Medicine and is supported by the Medical Scientist Training Program (5T32 GMP 7200).

³ Timothy Sullivan is supported by United States Public Health Service Grant 1 KO7 AI 00190.

⁴ Charles Parker is an Investigator, Howard Hughes Medical Institute.